Recombinant
RabMAb

Anti-Tryptophan Hydroxylase antibody [EP1311Y] (ab52954)

Overview

  • Product name
    Anti-Tryptophan Hydroxylase antibody [EP1311Y]
    See all Tryptophan Hydroxylase primary antibodies
  • Description
    Rabbit monoclonal [EP1311Y] to Tryptophan Hydroxylase
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide corresponding to residues near the C-terminus of human Tryptophan Hydroxylase

  • Positive control
    • THP-1 cell lysate and human urinary bladder carcinoma.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52954 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).
IP 1/50.
Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/50 - 1/100.

Target

  • Tissue specificity
    Isoform 2 seems to be less widely expressed than isoform 1.
  • Pathway
    Aromatic compound metabolism; serotonin biosynthesis; serotonin from L-tryptophan: step 1/2.
  • Sequence similarities
    Belongs to the biopterin-dependent aromatic amino acid hydroxylase family.
    Contains 1 ACT domain.
  • Information by UniProt
  • Database links
  • Alternative names
    • Indoleacetic acid 5 hydroxylase antibody
    • L tryptophan hydroxylase antibody
    • MGC119994 antibody
    • TPH 1 antibody
    • TPH antibody
    • TPH1 antibody
    • TPH1_HUMAN antibody
    • TPRH antibody
    • TRPH antibody
    • Tryptophan 5 hydroxylase 1 antibody
    • Tryptophan 5 monooxygenase 1 antibody
    • Tryptophan 5 monooxygenase antibody
    • Tryptophan 5-hydroxylase 1 antibody
    • Tryptophan 5-monooxygenase 1 antibody
    • Tryptophan hydroxylase 1 antibody
    see all

Images

  • Anti-Tryptophan Hydroxylase antibody [EP1311Y] (ab52954) at 1/500 dilution + THP-1 cell lysate at 10 µg

    Secondary
    goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 51 kDa
    Observed band size: 51 kDa

  • ab52954 at 1/50 dilution staining Tryptophan Hydroxylase in human urinary bladder carcinoma by Immunohistochemistry, Paraffin embedded tissue.
  • Overlay histogram showing HeLa cells stained with ab52954 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52954, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:
See all 14 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Jan 20 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Jan 08 2016

Question

1) Abcam product code ab 52954



2) Abcam order reference number or product batch number


3) Description of the problem weak staining



4) Sample preparation:

Species mouse

Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other: PFA/formalin fixed paraffin

Sample preparation

Positive control rat lung

Negative control igG



5) Fixation step

Yes

If yes: Fixative agent and concentration 10% neutral buffered formalin

Fixation time 24 hours

Fixation temperature room temp



6) Antigen retrieval method microwave 10mm citric acid buffer



7) Permeabilization method:

Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?no

Permeabilizing agent and concentration:





8) Blocking agent (eg BSA, serum…): bsa and normal horse serum

Concentration 10% bsa. 2.5% normal horse serum

Blocking time 1hour bsa . 30mins serum

Blocking temperature room temp



9) Endogenous peroxidases blocked?yes

Endogenous biotins blocked?no



10) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution 1:50 dilution

Diluent buffer 1% bsa in 50mm pbs

Incubation time overnight



11) Secondary antibody: ImmPress anti rabbit (vector labs)

Species:ready to use peroxidase linked secondary antibody

Reacts against:

Concentration or dilution prediluted

Diluent buffer

Incubation time

Fluorochrome or enzyme conjugate



12) Washing after primary and secondary antibodies:

Buffer tbs

Number of washes 4 x 15minutes



13) Detection method ImmPress VIP (vector labs)



14) How many times have you run this staining? Four times

Do you obtain the same results every time?yes

What steps have you altered to try and optimize the use of this antibody?

Exact same condition used with previous ab52954 with different batch number (please refer to my previous email)

Read More
Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will provide us with vital information for our monitoring of product quality. I would like to reassure you that this antibody is tested and covered by our guarantee for IHC.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there very few tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial.

I can recommend you may like to consider trying the following options:

1.) There is a difference between both vial concerning the concentration:

new lot: 1.5970 mg/ml
old lot : 2.8950 mg/ml

I therefore can suggest to increase the amount of antibody that is used. Our lab checked both lots in parallel and found no difference in staining when using both lot at a dilution of 1: 2000 in WB. The IHC was not checked again for the new lot.

2.) Could you confirm that all other reagents work as before? Is the secondary working?

Alternatively, or if these tips do not work, I am pleased to offer you a free of charge replacement or credit note in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Question
Answer

DISCOUNT CODE: ***
Expiration date: March 1, 2013
Value: $337

I am very pleased to hear you would like to accept our offer and test ab52954 with baboon samples. This code will give you $337 off your next order before the expiration date. To redeem this offer, please submit an Abreview for ab52954 with baboon samples and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active. For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/abtrial.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain, Raphe Nucleus)
Specification
Brain, Raphe Nucleus
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton X-100 0.1%
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 26 2012

Answer

Thank you for contacting us. The IHC-FrFl Abreview was submitted to us by one of our customer few months back. We had tried to troubleshoot the protocol however we were not able to resolve the issue. Because we do not have the IHC-Fr tested data of our own lab so we were unable to help customer with the problems they were experiencing. Generally the rabbit monoclonal are quite specific however in this case we were unable to find the cause of high background. We are also not sure whether a different protocol is needed or the antibody actually is unsuitable for IHC-FR. So I am sorry we will be unable to provide further information about its specificity in IHC-FR. We however also cannot judge the unsuitability of antibody from just one Abreview. As IHC-Fr is still untested application so I can provide an Abreview testing discount if you are interested in testing this ab again in IHC-Fr. The discount code will give your next antibody free of charge after submitting an IHC-Fr Abreview. We however also recommend using human bladder tissue sections as positive control. On the other hand we also have other antibody ab32821 which we have tested in IHC-Fr and is fully guaranteed to work in IHC-Fr with human or rat tissue sections. Other species are only predicted to react. Let me know if you would like to receive a discount code. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Rat Tissue sections (Hindbrain)
Specification
Hindbrain

Abcam user community

Verified customer

Submitted Apr 25 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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